Activated Protein C/TCF DNA decoy Redistributes β-catenin in Lipoplysaccharide-treated Human Aortic Endothelial Cells

• Main Authors: Chao,Tzupou, 趙梓棓
• Other Authors: Huang,Kuangtse
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/81401926306172967487

碩士 === 國立中正大學 === 化學工程研究所 === 100 === The patient with severe sepsis appears systemic inflammatory reaction, hypotension, prothrombotic diathesis, and organ dysfunction. In the previous study, activated protein C (APC) is effective to reduce the mortality from 30.8% to 24.7% in sepsis patients, However, the underlying mechanism of APC on the survival of lipopolysaccharide-activated endothelial cells (ECs) remains unclear. In our previous study, we found that APC is able to reduce the level of cytosolic β-catenin, so we further hypothesized that the effect of APC on cell survival and proinflammatory cytokine production may be due to inhibiting the β-catenin-TCF-mediated gene expression. To observe the dynamic concentration change of β-catenin in human aortic endothelial cells(HAECs) in response to lipopolysaccharide, APC, and DNA decoy, we transfected HAECs with a plasmid pEGFP-β-catenin, expressing β-catenin fusion protein with enhanced green fluorescent protein (EGFP). We observed that the fluorescence intensity in the cytosol showed insignificant change after HAECs treated with 0.1 g/mL lipopolysaccharide for one hour. For cells pretreated with lipopolysaccharide for six hours, the addition of APC or TCF DNA Decoy, decreased the cytosolic fluorescence intensity. These results suggest the efficacy of APC and TCF DNA decoy in reducing the amount of cytossolic β-catenin in HAECs.